Production and concentration of an antithyroid component



Patented Dec. 17, 1935 PRODUCTION AND dONCENTRATION OF AN ANTITHYROID COMPONENT Ferdinand Blum, Frankfort-on-the-Main, Germany, assignor to Siichsisches Serumwerk Aktiengesellschaft, Dresden, Germany No Drawing. Application June 4, 1932, Serial No.

4 Claims.

This invention relates to a method of obtaining and concentrating an antithyroid component. According to the invention, the antithyroid component is obtained from blood or organic tissue in so far as an antithyroid hormone is present or formed therein. I have found by experiments that every blood as well as organs, such as the spleen and liver, contain an antithyroid component which has an important influence on the regulation of the action of the thyroid gland.. This antithyroid effect is not due to the physical adsorption of thyroid gland substanceby the blood and, further,-does not depend on the colloidal condition of the blood but must nent of the blood. The antithyroid component which must be considered as counter-hormone (catechin) can be best characterized biologically 20 by saying that it will counteract or reduce the such as thyreoglobulin, thyroxin or the like, on amphibia (precipitated metamorphosis and stunted growth). The effects of certain amounts of thyroxin, for instance, whose converting power on tadpoles, for example, can be determined quantitatively, will be fully neutralized by accurately dosable amounts of the antithyroid component. Wherever, abnormally, thyroid iodine substance appears, the administration of anti- 30 hormone (catechin) is indicated.

The employment of the antithyroid hormone has been found to be of particular therapeutic value to act as protective matter against the phenomena of enlarged delivery of iodine albumen from the thyroid gland to the blood, as in Morbus Basedow.

Therapeutic employment of the antithyroid component has hitherto been handicapped by the fact that excessive amounts of the material containing the component had to be taken to produce a curative effect. A successful and efiective therapy was therefore impossible and it was'necossary to originate a method of obtaining the antithyroid component from the initial material,

freed from all unnecessary ballast, and of enrlching it.

The invention discloses a method of obtaining from the initial materials highly purified antithyroid hormone, so that one need not be satisfled with giving, as a maximum, 50 to 100 cu. cms. blood, but is in a position to give the eifective substance of say 2 liters of blood, 1. e., twenty to forty times the amount mentioned, concentrated in the form of 1 to 2 grams.-

dv c bebe atartibuted to a certain hormone-like compovery marked eifect of thyroid gland preparations,

In Germany June 17, 1931 comes still more remarkable when it is taken into consideration that it is possible to inject, at the highest, only a few cubic centimeters of the difllcultly sterilizable blood and that by the use of the foreign albumen this process involves always, 5 and particularly during repetition, grave dangers while the invention makes it possible to repeatedly inject the efiective component obtained from 1 to 2 liters of blood without any fear of injurious consequences.

The initial material is blood or organic tissue which contains the antithyroid component in larger quantities, such as the liver and spleen, which are the organs where the blood is formed or converted.

It has been ascertained that the antithyroid hormone is only loosely combined with albumen and can be separated therefrom by coagulation.

It is further soluble in water and in aqueous organic solvents, i. e., solvents miscible with water. The antithyroid hormone does not dialyze through a membrane.

The antithyroid component is obtained from the blood or its red corpuscles or the haemoglobin by separating the protein by coagulating agents.

The antithyroid component is thus freed from it's adherence to the protein and passes into the filtrate of the precipitation. A large portion of the antithyroid component, during the coagulation of the protein, passes immediately into the solution and the residual portion is extracted from the coagulation to which it adheres now in the free state by means of water or organic solvents soluble in water. Further purification of the extract enriched by antithyroid hormone by freeing it from salts and dialyzable impurities can be efiected by dialysis which can be applied also to the initial materials. Solvents, such as acetone or alcohol which are miscible with water,

may serve as coagulants, though coagulation by heat, possibly with the addition of suitable salts, metallic salts, colloidal iron and the like, will also free the antithyroid component from its adherence to protein.

The method mentionedabove would be carried out as follows:

Example 1.One liter of blood or of disintegrated tissue of liver or spleen is placed in a multiple quantity of acetone or alcohol whereby the protein constituents will be coagulated. After the solution'has been allowed to stand for some time the coagulation is separated from the solution, the solvent carefully distilled ofi and any .ifi-tjthfit has separated removed. The aqueous and other dialyzable substances.

solution is then concentrated or possibly dried and is then ready for use.

Additional quantities of effective antithyroid substances can be obtained from the coagulation by repeatedly treating the latter with one to two liters of cold or warm water or with aqueous alcohol or acetone. After separating the coagulation from the extract the solution is considerably concentrated. Dissolved protein can be precipitated with alcohol or acetone. The filtrate, on being freed from protein, is concentrated after driving out the solvent and can then be used.

Example 2.--Blo0d, red corpuscles'or haemoglobin are treated with low concentrations of acids whereupon the acids in the mixture are saturated by alkalies and the neutral solution is concentrated. The next step is to free this product from the protein by means of one of the coagulants stated, by adding for instance, alcohol, and

ing solution.

While the second part of the method is carried out as described in Example 2, the first part may, by way of example, be carried out as follows: One liter blood possibly after drying, is treated with one to two liters of sulfuric acid. From the extract the'sulfuric acid is precipitated by barium compounds" and the largest share of dissolved albumen is precipitated also. The neutral solution is concentrated and the protein which is still in solution precipitated according to Example 1.

Example 3.0ne liter of blood that has been rendered non-coagul'able is dialyzed 24 hours, once or twice, against water to free it from salts This precleaned blood may then be treated in accordance with Examples 1 and 2. Dialysis may further take place at any step of the processes described .in Examples 1 and 2 for instance after precipitating the protein.

I claim:--

1. A method of obtaining an antithyroid concentrate which comprises treating a substance of the class consisting of blood, liver and spleen to precipitate protein therefrom, filtering the precipitated protein, repeatedly extracting the precipitated proteinous matter with water to produce an aqueous extract, concentrating the filtrate and the aqueous extract'derived from the precipitate, and thereafter dialyzing and evaporating the filtrate and aqueous extract to dryness if necessary.

2. A method of obtaining an antithyroidconcentrate which comprises treating a substance of the class consisting of blood, liver and spleen with organic solvents to precipitate protein therefrom, filtering the precipitated protein, repeatedly extracting the precipitated proteinous matter with water to produce a substantially aqueous extract, concentrating the filtrate and the aqueous extract derived from the precipitate, and thereafter dialyzing and evaporating the filtrate and aqueous extract to dryness if necessary.

3. A method of obtaining. an antithyroid concentrate which comprises treating a substance oi the class consisting of blood, liver and spleen with heat to precipitate protein therefrom, filtering the precipitated protein, repeatedly extracting the precipitated proteinous matter with water to produce a substantially aqueous extract, concentrating the-filtrate and the aqueous extract derived from the precipitate, and thereafter dialyzing and evaporating the filtrate and aqueous extract to dryness if necessary.

4. A method of obtaining an antithyroid concentrate which comprises treating a substance of the class consisting of blood, liver and spleen with heat and acids to precipitate protein therefrom, filtering the precipitated protein, repeatedly extracting the precipitated proteinous matter with water to produce an aqueous extract, dialyzing and concentrating the filtrate and the aqueous extract derived from the precipitate, and evaporating the filtrate and aqueous extract to dryness if necessary.

FERDINAND BLUM. 

